Elsevier

Molecular Immunology

Volume 41, Issue 9, July 2004, Pages 873-884
Molecular Immunology

STAT-1 is activated by IL-4 and IL-13 in multiple cell types

https://doi.org/10.1016/j.molimm.2004.04.027Get rights and content

Abstract

While interleukin-4 (IL-4) and interleukin-13 (IL-13) can utilize a common receptor, composed of IL-4Rα and IL-13Rα1, IL-4 can also signal through a receptor with IL-4Rα and the common γchain (γC) as its subunits. IL-4 and IL-13 have been reported to elicit similar biological effects in a number of settings, including stimulating Ig isotype switching to IgE and inducing chemokines and cytokines in a variety of cell types whereas, depending on the receptor expression on responder cells, differential effects such as induction of type II helper T cell differentiation by IL-4 but not by IL-13 are also well documented. Recent data suggest distinct roles for these two cytokines in the ‘in vivo’ pathology of airway inflammatory diseases such as asthma. In this study, we examined the possibility of differential signaling by IL-4 and IL-13 on cells of the airway, by comparing expression of receptor chains and activation of different Signal Transducer and Activator of Transcription (STAT) family members. Five primary cultured cell lines representing four non-immune human lung tissue cell types (smooth muscle, epithelium, endothelium, and fibroblast) were utilized. While we readily detected expression of IL-4 Rα and IL-13Rα1 in all five cell lines, γC was not detectable in any of these cell lines. Consistent with previous reports, we detected STAT-6 activation in all five airway tissue cell lines examined in response to both cytokines. In addition, we also consistently detected STAT-1 activation in all of these cells. This observation was extended to include lymphoid as well as myeloid cells that express also γC chain. In conclusion, while the study found no differences in STAT activation in response to the two cytokines, the data show that in addition to STAT-6 activation, STAT-1 activation is also a part of the integral signaling pathways utilized by IL-4 and IL-13.

Introduction

Interleukin-4 (IL-4) and the closely related cytokine interleukin-13 (IL-13) share many immune regulatory functions, including enhanced expression of MHC class II and CD23 (FcεRII) genes (Chomarat and Banchereau, 1998, Jiang et al., 2000, Zurawski and de Vries, 1994), Ig class switching to IgE and IgG4 (Nelms et al., 1999), increased expression of adhesion molecules (Bochner et al., 1995, Doucet et al., 1998) and modulation of pro-inflammatory cytokine/chemokine production (Bonecchi et al., 2000, Deckers et al., 1998). Most of the activities of IL-4 and IL-13 can be ascribed to the activation of Signal Transducer and Activator of Transcription-6 (STAT-6) (Takeda et al., 1996). More recent studies, however, indicate that involvement from other signaling molecules, such as IRS-1/2, PI3 kinase and tyrosine phosphatases SHP-1 and SHP-2, are also crucial (Chomarat and Banchereau, 1998, Haque et al., 1998, Imani et al., 1997, Jiang et al., 2000, Keegan et al., 1994, Nelms et al., 1999, Wang et al., 1993, Wright et al., 1997).

Different types of IL-4/IL-13 receptors have been characterized (Aman et al., 1996, Donaldson et al., 1998, Keegan et al., 1994, Murata et al., 1998, Nelms et al., 1999, Orchansky et al., 1997, Zhang et al., 1997). Whereas the IL-4Rα chain and the common γ (γC) chain constitute the type I receptor, which is utilized solely by IL-4 (Keegan et al., 1994, Nelms et al., 1999), IL-4Rα and IL-13Rα1 form the type II receptor, a receptor shared by both IL-4 and IL-13 (Aman et al., 1996, Hilton et al., 1996). It has been well established that both human T cells and murine B cells do not express IL-13Rα1, precluding a response to IL-13 (Chomarat and Banchereau, 1998). Another high-affinity IL-13 binding receptor, IL-13Rα2, has been cloned from both human and mice, but no signaling capacity has yet been demonstrated (Donaldson et al., 1998, Zhang et al., 1997).

Both IL-4 and IL-13 are key modulators of immune responses in the asthmatic lung. These two cytokines exert their functions by recruiting lymphocytes and professional inflammatory cells, such as eosinophils and macrophages (D’Ambrosio et al., 2001), which contribute to airways hyperresponsiveness and, eventually, to remodeling of pulmonary structure (Holgate et al., 2000, Wills-Karp, 1999). Recent data utilizing animal models of allergic asthma indicate that IL-13 plays a central role in disease independent of IL-4 (Grunig et al., 1998, Wills-Karp et al., 1998). In an effort to determine if this is due to differences in signaling pathways employed by IL-4 and IL-13, we examined STAT family member activation by both cytokines in four different airway cell types. While we found no differences in STAT activation in response to IL-4 or IL-13, our results clearly indicate that, in addition to STAT-6 activation, STAT-1 is activated in response to both of these cytokines.

Section snippets

Cells, cytokines and antibodies

Bronchial smooth muscle cell (BSMC), human microvascular endothelial cell (HMVEC), normal human bronchial epithelial (NHBE), normal human lung fibroblast (NHLF), small airway epithelial cell (SAEC), normal human dermal fibroblast (NHDF), human mammary epithelial cell (HMEC) and human umbilical vein endothelial cell (HUVEC) cells are primary human cell lines purchased from Clonetics (San Diego, CA). They were cultured according to the instructions provided by the vendor. BSMC, HMVEC, NHBE, NHLF,

Expression of IL-4/IL-13 receptor subunits in primary human airway tissue cells

We first used ribonuclease protection assay (RPA) to examine the expression of all known IL-4/IL-13 receptor complex subunits in five primary human airway tissue cells: bronchial smooth muscle cells, human microvascular endothelial cells, normal human bronchial epithelial cells, normal human lung fibroblasts and small airway epithelial cells. A375, a natural IL-13Rα2-expressing cell line, and testis RNA were included as controls. As shown in Fig. 1, mRNAs for IL-4Rα and IL-13Rα1, components of

Discussion

Increasing evidence indicates that IL-13 plays a potent and unique role in the pathogenesis observed in asthma, parasitic infections and other pathological conditions. IL-4 shares many functional properties with IL-13, presumably because they share a common receptor composed of IL-4Rα and IL-13Rα1. In this study, we analyzed the direct effects of IL-4 and IL-13 on signal transduction in primary airway tissue cell lines by examining the activation of STAT proteins.

The specific activation of

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